Latex particle enhanced immunoturbidimetry based kit

All assays were performed by the Analytical Biochemistry Shared Resource at the University of Hawaii Cancer Center as reported for the original nested-case control studies following the manufacturer’s protocol unless noted otherwise [22 (link), 25 ]. Briefly, frozen serum samples from the MEC biorepository were analyzed in duplicate to quantify leptin and adiponectin using a double-antibody enzyme-linked-immunosorbent-assay (R&D Systems, Minneapolis, MN, U.S.A.). CRP was assessed using a Cobas MiraPlus clinical chemistry analyzer (Roche Diagnostics, Indianapolis, IN) and a latex particle enhanced immunoturbidimetry-based kit from Pointe Scientific (Lincoln Park, MI). TNF-α and IL-6 were included in the Luminex panel and were measured using a slight modification of an Invitrogen (Carlsbad, CA) magnetic high sensitivity 10-plex assay (LHC0001) and quantified on a Luminex 200 plate reader [22 (link), 25 ]. As reported previously [22 (link), 25 ], intra-batch coefficients of variation based on 96 blinded duplicate and 9 triplet samples for leptin, adiponectin, CRP, TNF-α, and IL-6 were 3.4 –6.4%, 2.5–9.4 %, 3.5–5.0%, 10.0%, and 8.9% respectively.

All assays were performed by the Analytical Biochemistry Shared Resource at the University of Hawaii Cancer Center as reported for the original nested-case control studies following the manufacturer's protocol unless noted otherwise.^{16 (link),19 (link)} Briefly, frozen serum samples were retrieved from the MEC biorepository and analyzed in duplicate to quantify leptin and adiponectin using double-antibody enzyme-linked-immunosorbent-assay (ELISA; R&D Systems, Minneapolis, MN, U.S.A.). CRP was assessed using a Cobas MiraPlus clinical chemistry analyzer (Roche Diagnostics, Indianapolis, IN) and a latex particle enhanced immunoturbidimetry based kit from Pointe Scientific (Lincoln Park, MI). The Luminex panel included IL-6 and TNF-α, which were measured using a slight modification of an Invitrogen (Carlsbad, CA) magnetic high sensitivity 10-plex assay (LHC0001) and Luminex 200 plate reader.^{16 (link),19 (link)} As reported previously^{16 (link),19 (link)} intra-batch coefficients of variation based on 96 blinded duplicate and 9 triplet samples for leptin, adiponectin, CRP, IL-6, and TNF-α were 3.4 -6.4%, 2.5-9.4 %, 3.5-5.0%, 8.9%, and 10.0%, respectively.